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Nat Methods. 2016 Dec;13(12):997-1000. doi: 10.1038/nmeth.4032. Epub 2016 Oct 17.

Genetic code expansion for multiprotein complex engineering.

Author information

1
European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
2
Karlsruhe Institute of Technology (KIT), Institute of Organic Chemistry, Karlsruhe, Germany.
3
European Molecular Biology Laboratory (EMBL), Grenoble, Grenoble, France.
4
Wellcome Trust Centre for Cell Biology, Institute of Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.
5
Vienna Biocenter Core Facilities (VBCF GmbH), Vienna, Austria.
6
Division of Pathology and Laboratory Medicine, European Institute of Oncology, Milan, Italy.
7
Chair of Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.
8
The School of Biochemistry, University of Bristol, Bristol, United Kingdom.
9
Karlsruhe Institute of Technology (KIT), Institut für Toxikologie und Genetik, Eggenstein-Leopoldshafen, Germany.

Abstract

We present a baculovirus-based protein engineering method that enables site-specific introduction of unique functionalities in a eukaryotic protein complex recombinantly produced in insect cells. We demonstrate the versatility of this efficient and robust protein production platform, 'MultiBacTAG', (i) for the fluorescent labeling of target proteins and biologics using click chemistries, (ii) for glycoengineering of antibodies, and (iii) for structure-function studies of novel eukaryotic complexes using single-molecule Förster resonance energy transfer as well as site-specific crosslinking strategies.

PMID:
27749839
DOI:
10.1038/nmeth.4032
[Indexed for MEDLINE]

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